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lipofectamine rnaimax  (Santa Cruz Biotechnology)


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    Structured Review

    Santa Cruz Biotechnology lipofectamine rnaimax
    Lipofectamine Rnaimax, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 6363 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lipofectamine+rnaimax/Control+siRNA-A/pm41595728-79-19-15
    Average 96 stars, based on 6363 article reviews
    lipofectamine rnaimax - by Bioz Stars, 2026-10
    96/100 stars

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    Related Articles

    Transfection:

    Article Title: Increased circulating miR-370-3p regulates steroidogenic factor 1 in endometriosis
    Article Snippet: .. Briefly, cells were transfected at 65% confluence in a six-well tissue culture plates with either SF-1 plasmid (no. sc-402334-ACT, 10 μg/ml) and hsa-miR-370-3p mimic together or siRNA plasmid (no. sc-402334, 1 μg/ml) later with hsa-miR-370-3p mimic using Lipofectamine RNAiMAX reagent according to the manufacturer’s protocol, with respective controls (no. sc-437275 control for overexpression of mimic; no. sc-418922 for the siRNA negative control) purchased from Santa Cruz Biotechnology (Dallas TX). ..

    Article Title: Acacetin Attenuates Lysophosphatidylcholine-Induced Vascular Smooth Muscle Cell Injury via Sirt1-Nrf2/p62 Signaling Axis.
    Article Snippet: .. For siRNA-mediated gene silencing experiments, cells transfected with Nrf2-targeting siRNA (50 nM) or scramble control (Santa Cruz, sc-37007) using Lipofectamine RNAiMAX were cultured for 48 h for protein knockdown before 24 h acacetin (3 μM) treatment. .. Following treatments, cells were loaded with 10 μM DCFH-DA in serum-free DMEM (37 ◦C, 30 min, dark), washed twice with Hanks’ Balanced Salt Solution (HBSS), and immediately analyzed by flow cytometry (Beckman Coulter Gallios, excitation/emission: 488/525 nm).

    Article Title: miR-222-3p is involved in neural tube closure by directly targeting Ddit4 in RA induced NTDs mouse model.
    Article Snippet: 5 × 105 cells/ ml was plated in antibiotic-free medium for 24 h before transfections. miR-222-3p mimics (mim222), miR-222-3p inhibitor (anti-222), miR mimics negative control (mim-NC), miR inhibitor negative control (anti-NC) and siRNA-Ddit4 plasmid were designed and synthesized by GenePharma Co (Shanghai, China). .. When the HT22 cells become 75–85% confluent, cells were transiently transfected with 100 nM miRNA or siRNA using Lipofectamine RNAiMax (Santa Cruz Biotechnology, USA), according to the manufacturer’s instructions. ..

    Article Title: The Reciprocal Causation of the ASK1-JNK1/2 Pathway and Endoplasmic Reticulum Stress in Diabetes-Induced Cognitive Decline
    Article Snippet: CCK8 assay (Beyotime, Shanghai, China) was used to detect the optimum concentration of HG. .. After seeding for 24 h, the cells were cultured in HG (100 mM) media with or without 2 mmol/L ER stress inhibitor 4-phenylbutyric acid (4-PBA) to inhibit ER stress for 24 h. Additionally, ASK1 small interfering RNA (ASK1-siRNA; sc-29749, Santa Cruz Biotechnology, CA, United States) and control siRNA (sc-37007) were used to inhibit ASK1 expression in cells for 24 h. Lipofectamine RNAiMAX (Santa Cruz Biotechnology) was used according to the manufacturer’s protocol for transfection of siRNA into the cells. ..

    Article Title: Thrombospondin-1 Receptor CD47 Overexpression Contributes to P-Glycoprotein-Mediated Multidrug Resistance Against Doxorubicin in Thyroid Carcinoma FTC-133 Cells
    Article Snippet: .. FTC-133R cells were transfected with non-targeting control scRNA (scRNA-cells) or siRNA specific to CD-47 (siRNA-cells) using Lipofectamine RNAiMAX following the Santa Cruz Biotechnology Inc (USA) the manufacturer’s instructions. ..

    Article Title: Acacetin Attenuates Lysophosphatidylcholine-Induced Vascular Smooth Muscle Cell Injury via Sirt1-Nrf2/p62 Signaling Axis
    Article Snippet: .. For siRNA-mediated gene silencing experiments, cells transfected with Nrf2-targeting siRNA (50 nM) or scramble control (Santa Cruz, sc-37007) using Lipofectamine RNAiMAX were cultured for 48 h for protein knockdown before 24 h acacetin (3 μM) treatment. .. Following treatments, cells were loaded with 10 μM DCFH-DA in serum-free DMEM (37 °C, 30 min, dark), washed twice with Hanks’ Balanced Salt Solution (HBSS), and immediately analyzed by flow cytometry (Beckman Coulter Gallios, excitation/emission: 488/525 nm).

    Article Title: The ANGPTL3-integrin α5 axis drives retinal vascular leakage in diabetic retinopathy.
    Article Snippet: The precleared lysates were incubated overnight at 4°C with gentle rotation in the presence of anti-ANGPTL3 (Thermo Fisher Scientific, Cat# MA5-35681, RRID: AB_2849581), antiintegrin α5 (Abcam, Cat# ab179475, RRID: AB_2716738), or antiIgG. .. The samples were separated by SDS-PAGE, and immunoprecipitated proteins were detected by immunoblotting using appropriate secondary antibodies. siRNA transfection HUVECs at 60% confluence were transfected with 50 nmol/L of integrin α5-specific siRNA (Santa Cruz Biotechnology, Cat# sc29372) or a scrambled control siRNA using Lipofectamine RNAiMAX .. AR TIC LE IN PR ES S ARTICLE IN PRESS 15 (Invitrogen, Cat# 11668-019) in accordance with the manufacturer’s protocol.

    Plasmid Preparation:

    Article Title: Increased circulating miR-370-3p regulates steroidogenic factor 1 in endometriosis
    Article Snippet: .. Briefly, cells were transfected at 65% confluence in a six-well tissue culture plates with either SF-1 plasmid (no. sc-402334-ACT, 10 μg/ml) and hsa-miR-370-3p mimic together or siRNA plasmid (no. sc-402334, 1 μg/ml) later with hsa-miR-370-3p mimic using Lipofectamine RNAiMAX reagent according to the manufacturer’s protocol, with respective controls (no. sc-437275 control for overexpression of mimic; no. sc-418922 for the siRNA negative control) purchased from Santa Cruz Biotechnology (Dallas TX). ..

    Article Title: Ethionine-mediated reduction of S -adenosylmethionine is responsible for the neural tube defects in the developing mouse embryo-mediated m6A modification and is involved in neural tube defects via modulating Wnt/β-catenin signaling pathway
    Article Snippet: The cells were treated with 20 mmol/L ethionine, 2 mmol/L SAM for 48 h. The siRNA-METTL3 plasmid was synthesized by Sangon Biotech (Shanghai, China). .. The METTL3 plasmid was synthesized by Gene Pharma (Shanghai, China), and transfected into the cells using Lipofectamine RNAiMax (Santa Cruz) according to the manufacturer’s instruction. ..

    Control:

    Article Title: Increased circulating miR-370-3p regulates steroidogenic factor 1 in endometriosis
    Article Snippet: .. Briefly, cells were transfected at 65% confluence in a six-well tissue culture plates with either SF-1 plasmid (no. sc-402334-ACT, 10 μg/ml) and hsa-miR-370-3p mimic together or siRNA plasmid (no. sc-402334, 1 μg/ml) later with hsa-miR-370-3p mimic using Lipofectamine RNAiMAX reagent according to the manufacturer’s protocol, with respective controls (no. sc-437275 control for overexpression of mimic; no. sc-418922 for the siRNA negative control) purchased from Santa Cruz Biotechnology (Dallas TX). ..

    Article Title: Acacetin Attenuates Lysophosphatidylcholine-Induced Vascular Smooth Muscle Cell Injury via Sirt1-Nrf2/p62 Signaling Axis.
    Article Snippet: .. For siRNA-mediated gene silencing experiments, cells transfected with Nrf2-targeting siRNA (50 nM) or scramble control (Santa Cruz, sc-37007) using Lipofectamine RNAiMAX were cultured for 48 h for protein knockdown before 24 h acacetin (3 μM) treatment. .. Following treatments, cells were loaded with 10 μM DCFH-DA in serum-free DMEM (37 ◦C, 30 min, dark), washed twice with Hanks’ Balanced Salt Solution (HBSS), and immediately analyzed by flow cytometry (Beckman Coulter Gallios, excitation/emission: 488/525 nm).

    Article Title: The Reciprocal Causation of the ASK1-JNK1/2 Pathway and Endoplasmic Reticulum Stress in Diabetes-Induced Cognitive Decline
    Article Snippet: CCK8 assay (Beyotime, Shanghai, China) was used to detect the optimum concentration of HG. .. After seeding for 24 h, the cells were cultured in HG (100 mM) media with or without 2 mmol/L ER stress inhibitor 4-phenylbutyric acid (4-PBA) to inhibit ER stress for 24 h. Additionally, ASK1 small interfering RNA (ASK1-siRNA; sc-29749, Santa Cruz Biotechnology, CA, United States) and control siRNA (sc-37007) were used to inhibit ASK1 expression in cells for 24 h. Lipofectamine RNAiMAX (Santa Cruz Biotechnology) was used according to the manufacturer’s protocol for transfection of siRNA into the cells. ..

    Article Title: Thrombospondin-1 Receptor CD47 Overexpression Contributes to P-Glycoprotein-Mediated Multidrug Resistance Against Doxorubicin in Thyroid Carcinoma FTC-133 Cells
    Article Snippet: .. FTC-133R cells were transfected with non-targeting control scRNA (scRNA-cells) or siRNA specific to CD-47 (siRNA-cells) using Lipofectamine RNAiMAX following the Santa Cruz Biotechnology Inc (USA) the manufacturer’s instructions. ..

    Article Title: Acacetin Attenuates Lysophosphatidylcholine-Induced Vascular Smooth Muscle Cell Injury via Sirt1-Nrf2/p62 Signaling Axis
    Article Snippet: .. For siRNA-mediated gene silencing experiments, cells transfected with Nrf2-targeting siRNA (50 nM) or scramble control (Santa Cruz, sc-37007) using Lipofectamine RNAiMAX were cultured for 48 h for protein knockdown before 24 h acacetin (3 μM) treatment. .. Following treatments, cells were loaded with 10 μM DCFH-DA in serum-free DMEM (37 °C, 30 min, dark), washed twice with Hanks’ Balanced Salt Solution (HBSS), and immediately analyzed by flow cytometry (Beckman Coulter Gallios, excitation/emission: 488/525 nm).

    Article Title: The ANGPTL3-integrin α5 axis drives retinal vascular leakage in diabetic retinopathy.
    Article Snippet: The precleared lysates were incubated overnight at 4°C with gentle rotation in the presence of anti-ANGPTL3 (Thermo Fisher Scientific, Cat# MA5-35681, RRID: AB_2849581), antiintegrin α5 (Abcam, Cat# ab179475, RRID: AB_2716738), or antiIgG. .. The samples were separated by SDS-PAGE, and immunoprecipitated proteins were detected by immunoblotting using appropriate secondary antibodies. siRNA transfection HUVECs at 60% confluence were transfected with 50 nmol/L of integrin α5-specific siRNA (Santa Cruz Biotechnology, Cat# sc29372) or a scrambled control siRNA using Lipofectamine RNAiMAX .. AR TIC LE IN PR ES S ARTICLE IN PRESS 15 (Invitrogen, Cat# 11668-019) in accordance with the manufacturer’s protocol.

    Over Expression:

    Article Title: Increased circulating miR-370-3p regulates steroidogenic factor 1 in endometriosis
    Article Snippet: .. Briefly, cells were transfected at 65% confluence in a six-well tissue culture plates with either SF-1 plasmid (no. sc-402334-ACT, 10 μg/ml) and hsa-miR-370-3p mimic together or siRNA plasmid (no. sc-402334, 1 μg/ml) later with hsa-miR-370-3p mimic using Lipofectamine RNAiMAX reagent according to the manufacturer’s protocol, with respective controls (no. sc-437275 control for overexpression of mimic; no. sc-418922 for the siRNA negative control) purchased from Santa Cruz Biotechnology (Dallas TX). ..

    Negative Control:

    Article Title: Increased circulating miR-370-3p regulates steroidogenic factor 1 in endometriosis
    Article Snippet: .. Briefly, cells were transfected at 65% confluence in a six-well tissue culture plates with either SF-1 plasmid (no. sc-402334-ACT, 10 μg/ml) and hsa-miR-370-3p mimic together or siRNA plasmid (no. sc-402334, 1 μg/ml) later with hsa-miR-370-3p mimic using Lipofectamine RNAiMAX reagent according to the manufacturer’s protocol, with respective controls (no. sc-437275 control for overexpression of mimic; no. sc-418922 for the siRNA negative control) purchased from Santa Cruz Biotechnology (Dallas TX). ..

    Cell Culture:

    Article Title: Acacetin Attenuates Lysophosphatidylcholine-Induced Vascular Smooth Muscle Cell Injury via Sirt1-Nrf2/p62 Signaling Axis.
    Article Snippet: .. For siRNA-mediated gene silencing experiments, cells transfected with Nrf2-targeting siRNA (50 nM) or scramble control (Santa Cruz, sc-37007) using Lipofectamine RNAiMAX were cultured for 48 h for protein knockdown before 24 h acacetin (3 μM) treatment. .. Following treatments, cells were loaded with 10 μM DCFH-DA in serum-free DMEM (37 ◦C, 30 min, dark), washed twice with Hanks’ Balanced Salt Solution (HBSS), and immediately analyzed by flow cytometry (Beckman Coulter Gallios, excitation/emission: 488/525 nm).

    Article Title: The Reciprocal Causation of the ASK1-JNK1/2 Pathway and Endoplasmic Reticulum Stress in Diabetes-Induced Cognitive Decline
    Article Snippet: CCK8 assay (Beyotime, Shanghai, China) was used to detect the optimum concentration of HG. .. After seeding for 24 h, the cells were cultured in HG (100 mM) media with or without 2 mmol/L ER stress inhibitor 4-phenylbutyric acid (4-PBA) to inhibit ER stress for 24 h. Additionally, ASK1 small interfering RNA (ASK1-siRNA; sc-29749, Santa Cruz Biotechnology, CA, United States) and control siRNA (sc-37007) were used to inhibit ASK1 expression in cells for 24 h. Lipofectamine RNAiMAX (Santa Cruz Biotechnology) was used according to the manufacturer’s protocol for transfection of siRNA into the cells. ..

    Article Title: Acacetin Attenuates Lysophosphatidylcholine-Induced Vascular Smooth Muscle Cell Injury via Sirt1-Nrf2/p62 Signaling Axis
    Article Snippet: .. For siRNA-mediated gene silencing experiments, cells transfected with Nrf2-targeting siRNA (50 nM) or scramble control (Santa Cruz, sc-37007) using Lipofectamine RNAiMAX were cultured for 48 h for protein knockdown before 24 h acacetin (3 μM) treatment. .. Following treatments, cells were loaded with 10 μM DCFH-DA in serum-free DMEM (37 °C, 30 min, dark), washed twice with Hanks’ Balanced Salt Solution (HBSS), and immediately analyzed by flow cytometry (Beckman Coulter Gallios, excitation/emission: 488/525 nm).

    Knockdown:

    Article Title: Acacetin Attenuates Lysophosphatidylcholine-Induced Vascular Smooth Muscle Cell Injury via Sirt1-Nrf2/p62 Signaling Axis.
    Article Snippet: .. For siRNA-mediated gene silencing experiments, cells transfected with Nrf2-targeting siRNA (50 nM) or scramble control (Santa Cruz, sc-37007) using Lipofectamine RNAiMAX were cultured for 48 h for protein knockdown before 24 h acacetin (3 μM) treatment. .. Following treatments, cells were loaded with 10 μM DCFH-DA in serum-free DMEM (37 ◦C, 30 min, dark), washed twice with Hanks’ Balanced Salt Solution (HBSS), and immediately analyzed by flow cytometry (Beckman Coulter Gallios, excitation/emission: 488/525 nm).

    Article Title: Acacetin Attenuates Lysophosphatidylcholine-Induced Vascular Smooth Muscle Cell Injury via Sirt1-Nrf2/p62 Signaling Axis
    Article Snippet: .. For siRNA-mediated gene silencing experiments, cells transfected with Nrf2-targeting siRNA (50 nM) or scramble control (Santa Cruz, sc-37007) using Lipofectamine RNAiMAX were cultured for 48 h for protein knockdown before 24 h acacetin (3 μM) treatment. .. Following treatments, cells were loaded with 10 μM DCFH-DA in serum-free DMEM (37 °C, 30 min, dark), washed twice with Hanks’ Balanced Salt Solution (HBSS), and immediately analyzed by flow cytometry (Beckman Coulter Gallios, excitation/emission: 488/525 nm).

    Synthesized:

    Article Title: Ethionine-mediated reduction of S -adenosylmethionine is responsible for the neural tube defects in the developing mouse embryo-mediated m6A modification and is involved in neural tube defects via modulating Wnt/β-catenin signaling pathway
    Article Snippet: The cells were treated with 20 mmol/L ethionine, 2 mmol/L SAM for 48 h. The siRNA-METTL3 plasmid was synthesized by Sangon Biotech (Shanghai, China). .. The METTL3 plasmid was synthesized by Gene Pharma (Shanghai, China), and transfected into the cells using Lipofectamine RNAiMax (Santa Cruz) according to the manufacturer’s instruction. ..

    Small Interfering RNA:

    Article Title: The Reciprocal Causation of the ASK1-JNK1/2 Pathway and Endoplasmic Reticulum Stress in Diabetes-Induced Cognitive Decline
    Article Snippet: CCK8 assay (Beyotime, Shanghai, China) was used to detect the optimum concentration of HG. .. After seeding for 24 h, the cells were cultured in HG (100 mM) media with or without 2 mmol/L ER stress inhibitor 4-phenylbutyric acid (4-PBA) to inhibit ER stress for 24 h. Additionally, ASK1 small interfering RNA (ASK1-siRNA; sc-29749, Santa Cruz Biotechnology, CA, United States) and control siRNA (sc-37007) were used to inhibit ASK1 expression in cells for 24 h. Lipofectamine RNAiMAX (Santa Cruz Biotechnology) was used according to the manufacturer’s protocol for transfection of siRNA into the cells. ..

    Expressing:

    Article Title: The Reciprocal Causation of the ASK1-JNK1/2 Pathway and Endoplasmic Reticulum Stress in Diabetes-Induced Cognitive Decline
    Article Snippet: CCK8 assay (Beyotime, Shanghai, China) was used to detect the optimum concentration of HG. .. After seeding for 24 h, the cells were cultured in HG (100 mM) media with or without 2 mmol/L ER stress inhibitor 4-phenylbutyric acid (4-PBA) to inhibit ER stress for 24 h. Additionally, ASK1 small interfering RNA (ASK1-siRNA; sc-29749, Santa Cruz Biotechnology, CA, United States) and control siRNA (sc-37007) were used to inhibit ASK1 expression in cells for 24 h. Lipofectamine RNAiMAX (Santa Cruz Biotechnology) was used according to the manufacturer’s protocol for transfection of siRNA into the cells. ..

    Immunoprecipitation:

    Article Title: The ANGPTL3-integrin α5 axis drives retinal vascular leakage in diabetic retinopathy.
    Article Snippet: The precleared lysates were incubated overnight at 4°C with gentle rotation in the presence of anti-ANGPTL3 (Thermo Fisher Scientific, Cat# MA5-35681, RRID: AB_2849581), antiintegrin α5 (Abcam, Cat# ab179475, RRID: AB_2716738), or antiIgG. .. The samples were separated by SDS-PAGE, and immunoprecipitated proteins were detected by immunoblotting using appropriate secondary antibodies. siRNA transfection HUVECs at 60% confluence were transfected with 50 nmol/L of integrin α5-specific siRNA (Santa Cruz Biotechnology, Cat# sc29372) or a scrambled control siRNA using Lipofectamine RNAiMAX .. AR TIC LE IN PR ES S ARTICLE IN PRESS 15 (Invitrogen, Cat# 11668-019) in accordance with the manufacturer’s protocol.

    Western Blot:

    Article Title: The ANGPTL3-integrin α5 axis drives retinal vascular leakage in diabetic retinopathy.
    Article Snippet: The precleared lysates were incubated overnight at 4°C with gentle rotation in the presence of anti-ANGPTL3 (Thermo Fisher Scientific, Cat# MA5-35681, RRID: AB_2849581), antiintegrin α5 (Abcam, Cat# ab179475, RRID: AB_2716738), or antiIgG. .. The samples were separated by SDS-PAGE, and immunoprecipitated proteins were detected by immunoblotting using appropriate secondary antibodies. siRNA transfection HUVECs at 60% confluence were transfected with 50 nmol/L of integrin α5-specific siRNA (Santa Cruz Biotechnology, Cat# sc29372) or a scrambled control siRNA using Lipofectamine RNAiMAX .. AR TIC LE IN PR ES S ARTICLE IN PRESS 15 (Invitrogen, Cat# 11668-019) in accordance with the manufacturer’s protocol.



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    mt-mKeima/YFP-Parkin HeLa cells were reverse transfected with siRNAs against the notated Rab GTPase family members for 24 h. Following <t>transfection,</t> cells were treated with vehicle or 30 μM FCCP for 4h to induce mitophagy. Cells were immediately live-cell imaged on a confocal microscope for mt-mKeima-based mitophagy analysis to analyze impact of Rab target knockdown on mitophagic flux. Results are graphed as average mt-mKeima (pH 4-5) positive foci per cell, normalized to respective plate scramble (Scr) siRNA controls. All plates also included an siRNA against PINK1 as a positive control for mitophagy impairment. Candidates with a ≥50% increase are presented in blue, and those with a ≥50% decrease in mitophagy readout are presented in red. n = 3 replicates (except Rab2B and Rab15: n = 2 replicates). Data are presented as mean ± SEM.
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    Image Search Results


    mt-mKeima/YFP-Parkin HeLa cells were reverse transfected with siRNAs against the notated Rab GTPase family members for 24 h. Following transfection, cells were treated with vehicle or 30 μM FCCP for 4h to induce mitophagy. Cells were immediately live-cell imaged on a confocal microscope for mt-mKeima-based mitophagy analysis to analyze impact of Rab target knockdown on mitophagic flux. Results are graphed as average mt-mKeima (pH 4-5) positive foci per cell, normalized to respective plate scramble (Scr) siRNA controls. All plates also included an siRNA against PINK1 as a positive control for mitophagy impairment. Candidates with a ≥50% increase are presented in blue, and those with a ≥50% decrease in mitophagy readout are presented in red. n = 3 replicates (except Rab2B and Rab15: n = 2 replicates). Data are presented as mean ± SEM.

    Journal: bioRxiv

    Article Title: Rab12 is a regulator of mitophagy and mitochondrial homeostasis

    doi: 10.64898/2026.03.29.715103

    Figure Lengend Snippet: mt-mKeima/YFP-Parkin HeLa cells were reverse transfected with siRNAs against the notated Rab GTPase family members for 24 h. Following transfection, cells were treated with vehicle or 30 μM FCCP for 4h to induce mitophagy. Cells were immediately live-cell imaged on a confocal microscope for mt-mKeima-based mitophagy analysis to analyze impact of Rab target knockdown on mitophagic flux. Results are graphed as average mt-mKeima (pH 4-5) positive foci per cell, normalized to respective plate scramble (Scr) siRNA controls. All plates also included an siRNA against PINK1 as a positive control for mitophagy impairment. Candidates with a ≥50% increase are presented in blue, and those with a ≥50% decrease in mitophagy readout are presented in red. n = 3 replicates (except Rab2B and Rab15: n = 2 replicates). Data are presented as mean ± SEM.

    Article Snippet: A working mix of Lipofectamine RNAiMAX transfection reagent (Fisher Scientific 13-778-150) and Opti-MEM reduced serum medium (Thermo Scientific 31985070) was prepared by diluting 300 μL RNAiMAX in 9.7 mL OptiMEM.

    Techniques: Transfection, Microscopy, Knockdown, Positive Control

    ( A ) RNA was collected for qRT-PCR analysis of Rab12 mRNA expression in mt-mKeima/YFP-parkin HeLa cells following 72 h transfection with siRNA against Rab12 (siRab12) or the scramble control (siScr). Internal control for normalization was GAPDH. **** p < 0.0001 determined by unpaired t-test. n = 3 biological replicates with three technical replicates each. ( B ) Representative western blot of lysates collected from mt-mKeima/YFP-parkin HeLa cells transfected with siScr or siRab12 assessed for Rab12 levels and β-actin as a loading control. ( C ) Quantification demonstrates Rab12 protein knockdown with Rab12 siRNA by the same protocol. **** p < 0.0001, as determined by unpaired t-test. n = 3 biological replicates. ( D ) Representative 60X confocal images of YFP-Parkin (green), mt-mKeima at neutral pH (cyan), and mt-mKeima at lysosomal pH (magenta) and ( E ) quantification of mt-mKeima mitophagy analysis demonstrated increased levels of mitophagy induced by treatment with 30 μM FCCP with Rab12 knockdown. ** p < 0.01, **** p < 0.0001, as determined by two-way ANOVA with Bonferroni’s multiple comparisons test. n = 3 biological replicates. Data are presented as mean ± SEM.

    Journal: bioRxiv

    Article Title: Rab12 is a regulator of mitophagy and mitochondrial homeostasis

    doi: 10.64898/2026.03.29.715103

    Figure Lengend Snippet: ( A ) RNA was collected for qRT-PCR analysis of Rab12 mRNA expression in mt-mKeima/YFP-parkin HeLa cells following 72 h transfection with siRNA against Rab12 (siRab12) or the scramble control (siScr). Internal control for normalization was GAPDH. **** p < 0.0001 determined by unpaired t-test. n = 3 biological replicates with three technical replicates each. ( B ) Representative western blot of lysates collected from mt-mKeima/YFP-parkin HeLa cells transfected with siScr or siRab12 assessed for Rab12 levels and β-actin as a loading control. ( C ) Quantification demonstrates Rab12 protein knockdown with Rab12 siRNA by the same protocol. **** p < 0.0001, as determined by unpaired t-test. n = 3 biological replicates. ( D ) Representative 60X confocal images of YFP-Parkin (green), mt-mKeima at neutral pH (cyan), and mt-mKeima at lysosomal pH (magenta) and ( E ) quantification of mt-mKeima mitophagy analysis demonstrated increased levels of mitophagy induced by treatment with 30 μM FCCP with Rab12 knockdown. ** p < 0.01, **** p < 0.0001, as determined by two-way ANOVA with Bonferroni’s multiple comparisons test. n = 3 biological replicates. Data are presented as mean ± SEM.

    Article Snippet: A working mix of Lipofectamine RNAiMAX transfection reagent (Fisher Scientific 13-778-150) and Opti-MEM reduced serum medium (Thermo Scientific 31985070) was prepared by diluting 300 μL RNAiMAX in 9.7 mL OptiMEM.

    Techniques: Quantitative RT-PCR, Expressing, Transfection, Control, Western Blot, Knockdown

    ( A ) mt-mKeima/YFP-Parkin HeLa were transfected with siRNA against Rab12 (siRab12) or a scramble control siRNA (siScr) for 72 h. Mito DNA DX analysis of mtDNA lesion frequency showed that Rab12 knockdown caused a significant decrease in mtDNA lesions relative to siScr transfection. n = 3 biological replicates. * p < 0.01, as determined by unpaired t-test. ( B ) No differences in steady state of mtDNA copy number was observed between conditions. n = 3 biological replicates. Data are presented as mean ± SEM.

    Journal: bioRxiv

    Article Title: Rab12 is a regulator of mitophagy and mitochondrial homeostasis

    doi: 10.64898/2026.03.29.715103

    Figure Lengend Snippet: ( A ) mt-mKeima/YFP-Parkin HeLa were transfected with siRNA against Rab12 (siRab12) or a scramble control siRNA (siScr) for 72 h. Mito DNA DX analysis of mtDNA lesion frequency showed that Rab12 knockdown caused a significant decrease in mtDNA lesions relative to siScr transfection. n = 3 biological replicates. * p < 0.01, as determined by unpaired t-test. ( B ) No differences in steady state of mtDNA copy number was observed between conditions. n = 3 biological replicates. Data are presented as mean ± SEM.

    Article Snippet: A working mix of Lipofectamine RNAiMAX transfection reagent (Fisher Scientific 13-778-150) and Opti-MEM reduced serum medium (Thermo Scientific 31985070) was prepared by diluting 300 μL RNAiMAX in 9.7 mL OptiMEM.

    Techniques: Transfection, Control, Knockdown